Spiro-substituted azacycles as neurokinin-3 antagonists

ABSTRACT

Disclosed are spiro-substituted azacycles of formula I   &lt;IMAGE&gt;  are selective neurokinin-3 antagonists useful in the treatment of CNS disorders.

BACKGROUND OF THE INVENTION

The invention disclosed herein is directed to certain spiro-substitutedazacycles useful as tachykinin receptor antagonists. In particular, thecompounds disclosed herein are neurokinin-3 receptor antagonists.

The tachykinins, substance P (SP), neurokinin A (NKA) and neurokinin B(NKB), are structurally similar members of a family of neuropeptides.Each of these is an agonist of the receptor types, neurokinin-1 receptor(NK-1), neuorokinin-2 receptor (NK-2) and neuorokinin-3 receptor (NK-3),which are so defined according to their unique amino acid sequence andtheir relative abilities to bind tachykinins with high affinity and tobe activated by the natural agonists SP, NKA and NKB respectively.

The tachykinins are distinguished by a conserved carboxyl-terminalsequence Phe-X-Gly-Leu-Met-NH₂. More specifically, substance P is apharmacologically-active neuropeptide that is produced in mammals andpossesses a characteristic amino acid sequence:

SEQ ID NO:1: Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH₂

Neurokinin A possesses the following amino acid sequence:

SEQ ID NO:2: His-Lys-Thr-Asp-Ser-Phe-Val-Gly-Leu-Met-NH₂.

Neurokmin B possesses the following amino acid sequence:

SEQ ID NO:3: Asp-Met-His-Asp-Phe-Phe-Val-Gly-Leu-Met-NH₂.

(Chang et al., Nature New Biol. 232, 86 (1971 ); D. F. Veber et al.,U.S. Pat. No. 4,680,283).

The neurokinin receptors are widely distributed throughout the mammaliannervous system (especially brain and spinal ganglia), the circulatorysystem and peripheral tissues (especially the duodenum and jejunum) andare involved in regulating a number of diverse biological processes.This includes sensory perception of olfaction, vision, audition andpain, movement control, gastric motility, vasodilation, salivation, andmicturition (B. Pernow, Pharmacol. Rev., 1983, 35, 85-141). The NK1 andNK2 receptor subtypes are implicated in synaptic transmission(Laneuville et al., Life Sci., 42:1295-1305 (1988)).

Substance P acts as a vasodilator, a depressant, stimulates salivationand produces increased capillary permeability. It is also capable ofproducing both analgesia and hyperalgesia in animals, depending on doseand pain responsiveness of the animal (see R. C. A. Frederickson et al.,Science, 199, 1359 (1978); P. Oehme et al., Science, 208, 305 (1980))and plays a role in sensory transmission and pain perception (T. M.Jessell, Advan. Biochem. Psychopharmacol. 28, 189 (1981 )). Inparticular, substance P has been shown to be involved in thetransmission of pain in migraine (see B. E. B. Sandberg et al., Journalof Medicinal Chemistry, 25, 1009 (1982)), and in arthritis (Levine etal. Science, (1984) 226 547-549).

In the airways, it has been indicated that NK1 receptors are associatedwith microvascular leakage and mucus secretion, while NK2 receptorsregulate smooth muscle contraction. Also, it has been shown that bothsubstance P and neurokinin A are effective in inducing airwayconstriction and edema. Based on such findings, it is believed thatsubstance P and neurokinin A may be involved in the pathogenesis ofneurogenic inflammation, including allergic diseases such as asthma.(Frossard et al., Life Sci., 49, 1941-1953 (1991); Advenier, et al.,Biochem. Biophys. Res. Comm., 184(3), 1418-1424 (1992)).

In experimental studies, sensory neuropeptides, especially tachykininssuch as substance P and neurokinin A, can bring about many of thepathophysiological features of asthma. Neurokinin A is a very potentconstrictor of human airways in vitro, and substance P causes mucussecretion in the airways. (Barnes P. J., Lancet, pp242-44 (1986); RogersD. R., Aursudkij B., Barnes P. J., Euro. J. Pharmacol, 174, 283-86(1989)).

Inhalation of bradykinin causes bronchoconstriction in asthmaticpatients but not in normal subjects. (Fuller R. W., Dixon C. M. S., CussF. M. C., Barnes P. J., Am Rev Respir Dis, 135, 176-80 (1987)). Sincethe bradykinin-induced bronchoconstriction is partly opposed byanticholinergic agents and since bradykinin is only a weak constrictorof human airways in vitro, it has been suggested that thebronchoconstrictor response is partly mediated by a neural reflex.Bradykinin stimulates vagal afferent C fibers and causesbronchoconstriction in dogs. (Kaufman M. P., Coleridge H. M., ColeridgeJ. C. G., Baker D. G., J. Appl. Physio., 48, 511-17 (1980)). Inguinea-pig airways, bradykinin causes a bronchoconstrictor response byway of cholinergic and sensory-nerve-mediated mechanisms. (Ichinoe M.,Belvisi M. G., Barnes P. J., J. Pharmacol. Exp. Ther., 253, 594-99(1990). Bradykinin-induced bronchoconstriction in human airways maytherefore be due partly to tachykinin released from sensory nerveterminals via axon reflex mechanisms. Clinical trials have shown that adual NK-1/NK-2 antagonist (such as FK-224) protects against bradykinininduced bronchocontriction in asthmatic patients. (Ichinoe, M. et al.,Lancet,, vol. 340, pp 1248-1251 (1992)).

The tachykinins have also been implicated in gastrointestinal (GI)disorders and diseases of the GI tract, such as inflammatory boweldisease, ulcerative colitis and Crohn's disease, etc. (see Mantyh etal., Neuroscience, 25 (3), 817-37 (1988) and D. Regoli in "Trends inCluster Headache" Ed. F. Sicuteri et al., Elsevier ScientificPublishers, Amsterdam, 1987, pp. 85-95).

It is also hypothesized that there is a neurogenic mechanism forarthritis in which substance P may play a role (Kidd et al., "ANeurogenic Mechanism for Symmetric Arthritis" in The Lancet, 11 November1989 and Gronblad et al., "Neuropeptides in Synovium of Patients withRheumatoid Arthritis and Osteoarthritis" in J. Rheumatol. (1988) 15(12)1807-10). Therefore, substance P is believed to be involved in theinflammatory response in diseases such as rheumatoid arthritis andosteoarthritis (O'Byrne et al., in Arthritis and Rheumatism (1990) 331023-8). Other disease areas where tachykinin antagonists are believedto be useful are allergic conditions (Hamelet et al., Can. J. Pharmacol.Physiol. (1988) 66 1361-7), immunoregulation (Lotz et al., Science(1988) 241 1218-21, Kimball et al., J. Immunol. (1988) 141 (10) 3564-9and A. Perianin, et al., Biochem. Biophys. Res. Commun. 161,520 (1989))vasodilation, bronchospasm, reflex or neuronal control of the viscera(Mantyh et al., PNAS (1988) 85 3235-9) and, possibly by arresting orslowing β-amyloid-mediated neurodegenerative changes (Yankner et al.,Science, (1990) 250, 279-82) in senile dementia of the Alzheimer type,Alzheimer's disease and Downs Syndrome. Substance P may also play a rolein demyelinating diseases such as multiple sclerosis and amyotrophiclateral sclerosis [J. Luber-Narod et. al., poster presented at C.I.N.P.XVIIIth Congress, 28th June-2nd July, 1992]. Antagonists selective forthe substance P and/or the neurokinin A receptor may be useful in thetreatment of asthmatic disease (Frossard et al., Life Sci., 49,1941-1953 (1991); Advenier, et al., Biochem. Biophys. Res. Comm.,184(3), 1418-1424 (1992)). These antagonists may also be useful in thetreatment of emesis. See C. Bountra, K. Bounce, T. Dale, C. Gardner, C.Jordan. D. Twissell and P. Ward, Eur. J. Pharmacol., 249. R3-R4 (1993)"Anti-emetic profile of a non-peptide neurokinin NK1 receptorantagonist, CP-99,994, in the ferret.

The localisation of tachykinins and neurokinin receptor subtypes withinthe striatum is also heterogeneous. NKB immunoreactive fibres arecolocalised within GABA containing neurones that project to thepalladium but not the substantia nigra pars reticulata, whereas the SPcontaining neurones project principally to the substantia nigra parsreticulata. See Burgunder, J. M., & Young, W. S. 1989. Distribution,projection, and dopaminergic regulation of the neurokinin BmRNA-containing neurones of the rat caudate-putamen. Neurosci. 32,323-335. Activation of tachykinin receptors in the straitum modulatesthe release of neurotransmitters including acetylcholine and dopamineSee Tremblay, L., Kemel, M-L. Desban, M., Gauchy, C., & Glowinski, J.1992. Distinct presynaptic control of dopamine release instirosomal-matrix-enriched areas of the rat striatum by selectiveagonists of NK1, NK2 and NK3 tachykinin receptors. Proc. Natl. Acad.Sci. U.S.A. 89, 11214-11218. Interestingly in that study the release ofdopamine by [Pro⁷ ]NKB (NK3) in the matrix compartment was insensitiveto tetrodotoxin suggesting a presynaptic localisation of NK₃ receptors.

This hypothesis is further supported by the finding that the NKB-inducedstimulation of acetylchlorine release in rat striatum is reduced by bothTTX and by lesions of the nigrostriatal pathway, and is consistent withthe presence of NK₃ receptors on dopamine cell bodies of thestriatonigral and mesolimbic pathways. See Arenas, E., Alberch, J.,Perez-Navarro, E., Solsona, C., Marsal, J. 1991. Neurokinin receptorsdifferentially mediate endogenous acetylcholine release evoked bytachykinins in the neostriatum. J. Neurosci. 11 (8), 2332-2338; andKeegan, K. D., Woodruff, G., & Pinnock, R. D. 1992. The selective NK3receptor agonist senktide excites a subpopulation of dopamine-sensitiveneurones inthe rat substantia nigra pars compact in vitro. Br. J.Pharmacol. 105, 3-5.

We have found that Tachykinin receptor subtype on presumed dopamineneurones of the rat ventral tegmental area are a NK3 and not a NK1 orNK2 receptor subtype. These data suggest that NK3 receptors mediate theprincipal excitatory influence of tachykinins on mesolimbic dopamineneurones; however the role of receptors on afferent projections to theVTA and the relative tone of neuropeptide-containing fibres may have amore significant influence over their function.

Binding studies have shown NK₃ -receptors to be present in brain slicesfrom several species eg rat, mouse and guinea-pig, however, Dietl &Palacios, using ¹²⁵ I-labelled Bolton Hunter (BH) eledoisin reported anabsence of NK3-receptors in primate and human brain. The humanNK3-receptor was cloned from human brain mRNA, indicating that theprotein is expressed in this tissue. See Huang, et al, BBRC 184:996-972(1992). However, the cloned human NK3-receptor has lower affinity foreledoisin thatn the rat receptor and this probably explains the apparentabsence of NK3-binding sites in human brain when 125I-BHeledoisin isused as the ligand. Indeed, using 3H-senktide Guard & Watson readilydemonstrated the presence of NK3-binding sites in primate brain. SeeDietl, M. M. & Palacios, J. M. Phylogeny and tachykinin receptorlocalisation in the vertebrate central nervous system: apparent absenceof neurokinin-2 and neurokinin-3 binding sites in the human brain. 1991Br Res 539:211-222; Buell, G., Schultz, S. J., Arkinstal, S. J., Maury,K., Missotten, M., Adami, N., Talabot, F. & Kawashin, E. Molecularcharacterisation, expression and localisation of human neurokinin-3receptor. 1992 Febs Letts 299, 90-95; and Guard, S. & Watson, S. P. 1991Neurochem Int 18:149-165.

Interestingly, infusion of senktide (an NK3-receptor agonist) bymicrodialysis in the substantia nigra and VTA of the rat causedbehavioural responses characteristic of the activation of dopaminergicpathways and this effect was different with age. This observationimplies that neurokinin receptors may play a role in centraldopaminergic disorders, particularly those such as Parkinsonism whichare more prelevant in advanced age. See Stoessl, A. J., Polanski, E. &Frydryszak, H. Effects of ageing on tachykinin function in the basalganglia. 1993 Brain Res 632: 21-28.

SUMMARY OF THE INVENTION

This invention is directed to compounds of formula I. ##STR2##

The invention is also concerned with pharmaceutical formulations withthese novel compounds as active ingredients and the use of the novelcompounds and their formulations in the treatment of certain disordersincluding CNS disorders.

The compounds of this invention are tachykinin receptor antagonists, inparticular, neurokinin-3 receptor antagonists, and are useful in thetreatment of inflammatory diseases, pain, migraine, asthma, emesis andCNS disorders.

DETAILED DESCRIPTION OF THE INVENTION

This invention is directed to compounds of formula I. ##STR3## or apharmaceutically acceptable salt thereof, wherein the nitrogen expresslyshown above is optionally quaternized with C₁₋₄ alkyl or phenyl C₁₋₄alkyl or is optionally present as the N-oxide (N⁺ O⁻), and wherein:

l and m are each independently 0, 1, 2, 3, 4, or 5, with the provisothat l+m is equal to 1, 2, 3, 4, or 5;

R₁ is ##STR4## M is selected from a group consisting of: linear orbranched C₁₋₈ alkyl, linear or branched C₂₋₈ alkenyl, wherein the C₁₋₈alkyl or C₂₋₈ alkenyl is optionally mono or di- substituted, thesubstitutents independently selected from:

(a) hydroxy,

(b) cyano,

(c) halogen which is defined to include Br, Cl, I, and F,

(d) trifluoromethyl,

(e) phenyl or mono, di or trisubstituted phenyl, the substitutentsindependently selected from

(1) phenyl,

(2) C₁₋₃ alkyl,

(3) C₁₋₃ alkoxy,

(4) cyano,

(5) halogen,

(6) trifluoromethyl,

(7) --NR₆ COR₇, wherein R₆ and R₇ are independently selected from:

(a) hydrogen,

(b) C₁₋₆ alkyl, or mono or disubstituted C₁₋₆ alkyl, the substitutentsindependently selected from

(1) phenyl,

(2) hydroxy,

(3) oxo,

(4) cyano,

(5) C₁₋₃ alkoxy,

(6) trifluoromethyl,

(c) phenyl or mono, di or trisubstituted phenyl, the substitutentsindependently selected from

(1) hydroxy,

(2) C₁₋₃ alkyl,

(3) cyano,

(4) halogen,

(5) trifluoromethyl,

or

R₆ and R₇ are joined together to form a 5-, 6-, or 7-membered monocyclicsaturated ring containing 1 or 2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, and in which the ring is unsubstituted ormono or disubstituted, the substituents independently selected from

(a) hydroxy,

(b) oxo,

(c) cyano,

(d) halogen,

(e) trifiuoromethyl,

(8) --NR₆ CO₂ R₇,

(9) --NR₆ CONHR₇,

(10) --NR₆ S(O)_(j) R₇, wherein j is 1 or 2,

(11 ) --CONR₆ R₇,

(12) --COR₆,

(13) --CO₂ R₆,

(14) --OR₆,

(15) --S(O)_(k) 'R₆ wherein k' is 0, 1 or 2,

(f) heteroaryl, wherein heteroaryl is selected from the group consistingof:

(1) benzimidazolyl,

(2) benzofuranyl,

(3) benzoxazolyl,

(4) furanyl,

(5) imidazolyl,

(6) indolyl,

(7) isooxazolyl,

(8) isothiazolyl,

(9) oxadiazolyl,

(10) oxazolyl,

(11) pyrazinyl,

(12) pyrazolyl,

(13) pyridyl,

(14) pyrimidyl,

(15) pyrrolyl,

(16) quinolyl,

(17) tetrazolyl,

(18) thiadiazolyl,

(19) thiazolyl,

(20) thienyl,

(21) triazolyl,

wherein the heteroaryl is unsubstituted or mono di or trisubstituted,the substituents independently selected from

(1) phenyl,

(2) hydroxy,

(3) cyano,

(4) halogen,

(5) trifluoromethyl,

Rb is selected from the group consisting of

(a) C₁₋₃ alkyl,

(b) hydroxyC₁₋₃ alkyl,

(c) hydrogen, and

(d) trifluoromethyl,

Rc is substituted branched or linear C₁₋₄ alkyl wherein the substituentis selected from the group consisting of

(a) C₁₋₃ alkyl,

(b) hydroxyC₁₋₃ alkyl

(c) C₁₋₃ alkoxy,

(d) halogen,

(e) trifluoromethyl;

(f) hydrogen,

(g) phenyl or mono, di or trisubstituted phenyl, the substitutentsindependently selected from

(1) hydroxy,

(2) cyano,

(3) halogen,

(4) trifluoromethyl,

(5) C₁₋₃ alkyl,

(6) C₁₋₃ alkoxy,

X is ##STR5## Rd is selected from a group consisting of: (1) hydrogen,

(2) C₁₋₈ linear or branched alkyl, unsubstituted, monosubstituted ormultiply substituted with

(a) --OH,

(b) --OR₆,

(c) ═O,

(d) --NHCOR₆,

(e) --NR₆ R₇,

(f) --CN,

(g) --halogen,

(h) --CF₃,

(i) --phenyl, unsubstituted or substituted, wherein the substitutentsare selected from the group consisting of

(1) hydroxy,

(2) cyano,

(3) halogen,

(4) trifluoromethyl,

(3) phenyl or mono, di or trisubstituted phenyl, the substituentsindependently selected from

(1) hydroxy,

(2) cyano,

(3) --C(O)NR₆ R₇,

(4) --NR₆ R₇,

(5) --NR₆ COR₇,

(6) --halogen,

(7) --CF₃, and

(8) C₁₋₃ alkyl,

(4) aryl or mono, di or trisubstituted aryl wherein the aryl is selectedfrom the group consisting of

(a) benzimidazolyl,

(b) benzofuranyl,

(c) benzoxazolyl,

(d) furanyl,

(e) imidazolyl,

(f) indolyl,

(g) isoxazolyl,

(h) isothiazolyl,

(i) oxadiazolyl,

(j) oxazolyl,

(k) pyrazinyl,

(l) pyrazolyl,

(m) pyridyl,

(n) pyrimidyl,

(o) pyrrolyl,

(p) quinolyl,

(q) tetrazolyl,

(r) thiadiazolyl,

(s) thiazolyl,

(t) thienyl, and

(u) triazolyl,

wherein the substituents are each independently selected from

(1) hydroxy,

(2) cyano,

(3) --C(O)NR₆ R₇,

(4) --NR₆ R₇,

(5) --NR₆ COR₇,

(6) --halogen,

(7) --CF₃, and

(8) C₁₋₃ alkyl,

Re is selected from the group consisting of

(1) C₁₋₆ alkyl, branched or unbranched, unsubstituted or mono ordisubstituted, the substituents being selected from hydrogen andhydroxy,

(2) hydroxy,

(3) OR₆, wherein R₆ is as defined immediately above,

(4) halogen,

(5) trifluoromethyl,

(6) nitro,

(7) cyano,

(8) NR₆ R₇,

(9) NHCOR₆,

(10) NR₆ COR₇,

(11) NHCO₂ R₆,

(12) NR₆ CO₂ R₇,

(13) NHS(O)_(j) R₆,

(14) NR₆ S(O)_(j) R₇,

(15) CONR₆ R₇,

(16) COR₆,

(17) CO₂ R₆, and

(18) S(O)_(j) R₆ ;

or Rd and Re are joined together to form a mono- or di- substitutedsaturated or unsaturated monocyclic or bicyclic ring of 5, 6, 7, 8, 9,or 10 atoms including the carbon to which Rd and Re are attached,wherein 0, 1, or 2 of said atoms may be heteroatoms independentlyselected from N, O, or S,

wherein the substituents are each independently selected from

(1) hydroxy,

(2) oxo,

(3) cyano,

(4) --C(O)NR₆ R₇,

(5) --NR₆ R₇,

(6) --NR₆ COR₇,

(7) --halogen,

(8) --CF₃,

(9) --C₁₋₃ alkyl,

(10) --S(O)2 C₁₋₃ alkyl, and

(11) --C(O)R₆.

One genus within this embodiment is the compounds of formula I wherein:

l+m is equal to 2, or 3;

R₁ is ##STR6## M is selected from a group consisting of: C₂, C₃ or C₄linear or branched alkyl, unsubstituted or mono, di or tri substituted,the substitutents independently selected from:

(a) hydroxy,

(b) Cl or F,

(c) phenyl or mono, di or trisubstituted phenyl, the substitutentsindependently selected from

(1) hydroxy,

(2) C₁₋₃ alkyl,

(3) halogen,

(4) --NR₆ COR₇, wherein R₆ and R₇ are independently selected from:

(a) hydrogen,

(b) C₁₋₄ alkyl, or mono or disubstituted C₁₋₄ alkyl, the substitutentsindependently selected from

(1) phenyl,

(2) hydroxy,

(3) oxo,

(4) halogen, or

(c) phenyl or mono di or trisubstituted phenyl, the substitutentsindependently selected from

(1) hydroxy,

(2) C₁₋₃ alkyl,

(3) cyano,

(4) halogen,

(5) trifluoromethyl,

or

R₆ and R₇ are joined together to form a 5-, 6-, or 7-membered monocyclicsaturated ting containing 1 or 2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur, and in which the ring is unsubstituted ormono or disubstituted, the substituents independently selected from

(a) hydroxy,

(b) oxo,

(c) cyano,

(d) halogen,

(5) --NR₆ CO₂ R₇,

(6) --NR₆ CONHR₇,

(7) --NR₆ S(O)_(j) R₇, wherein j is 1 or 2,

(8) --CONR₆ R₇,

(9) --COR₆,

(10) --CO₂ R₆,

(11) --OR₆,

(12) --S(O)_(k) 'R₆, wherein k' is 0, 1, or 2,

(d) heteroaryl, wherein heteroaryl is selected from the group consistingof:

(1) pyrazinyl,

(2) pyrazolyl,

(3) pyridyl,

(4) pyrimidyl,

(5) thienyl, and

(6) indolyl,

wherein the heteroaryl is unsubstituted or mono di or trisubstituted,the substituents independently selected from

(a) phenyl,

(b) hydroxy, and

(c) halogen,

Rb is selected from the group consisting of

(a) C₁₋₃ alkyl, and

(b) hydrogen,

Rc is substituted branched or linear C₁₋₄ alkyl wherein the substituentis selected from the group consisting of

(a) C₁₋₃ alkyl,

(b) C₁₋₃ alkoxy,

(c) hydrogen,

(d) phenyl or mono or di- isubstituted phenyl, the substitutentsindependently selected from

(1) halogen,

(2) trifluoromethyl,

(3) C₁₋₃ alkyl,

X is ##STR7## Rd is selected from a group consisting of: (1) C₁₋₈ linearor branched alkyl,unsubstituted, monosubstituted or multiply substitutedwith

(a) --OH,

(b) --OR₆,

(c) ═O,

(d) --NHCOR₆,

(e) --NR₆ R₇,

(f) --phenyl, unsubstituted or substituted, wherein the substitutentsare selected from the group consisting of

(1) hydroxy,

(2) cyano,

(3) halogen,

(4) trifluoromethyl,

(2) phenyl or mono or di- substituted phenyl, the substituentsindependently selected from

(1) hydroxy,

(2) cyano,

(3) --C(O)NR₆ R₇,

(4) --NR₆ R₇,

(5) --NR₆ COR₇,

(6) --halogen,

(7) --CF₃, and

(8) C₁₋₃ alkyl,

(3) aryl or mono, di or trisubstimted aryl wherein the aryl is selectedfrom the group consisting of

(a) benzimidazolyl,

(b) benzofuranyl,

(c) benzoxazolyl,

(d) furanyl,

(e) imidazolyl,

(f) indolyl,

(g) isooxazolyl,

(h) isothiazolyl,

(i) oxadiazolyl,

(j) oxazolyl,

(k) pyrazinyl,

(l) pyrazolyl,

(m) pyridyl,

(n) pyrimidyl,

(o) pyrrolyl,

(p) quinolyl,

(q) tetrazolyl,

(r) thiadiazolyl,

(s) thiazolyl,

(t) thienyl, and

(u) triazolyl,

wherein the substituents are independently selected from

(1) hydroxy,

(2) cyano,

(3) --NR₆ R₇,

(4) --NR₆ COR₇,

(5) --halogen,

(6) --CF₃, and

(7) C₁₋₃ alkyl,

Re is selected from the group consisting of

(1) C₁₋₆ alkyl, branched or unbranched, unsubstituted or mono ordisubstituted, the substituents being selected from hydrogen andhydroxy,

(2) hydroxy,

(3) OR₆, wherein R₆ is as defined immediately above,

(4) halogen,

(5) trifluoromethyl,

(6) nitro,

(7) cyano,

(8) NR₆ R₇,

(9) NHCOR₆,

(10) NR₆ COR₇,

(11) NHCO₂ R₆,

(12) NR₆ CO₂ R₇,

(13) NHS(O)_(j) R₆,

(14) NR₆ S(O)_(j) R₇,

(15) CONR₆ R₇,

(16) COR₆,

(17) CO₂ R₆, and

(18) S(O)_(j) R₆ ;

or R d and Re are joined together to form a mono- or di- substitutedsaturated or unsaturated monocyclic or bicyclic ring of 5, 6, 7, 8, 9,or 10 atoms including the carbon to which Rd and Re are attached,whrerin 0, 1, or 2 of said atoms may be heteroatoms independentlyselected from N, O, or S,

wherein the substituents independently selected from

(1) oxo,

(2) --C(O)NR₆ R₇,

(3) --NR₆ R₇,

(4) --NR₆ COR₇,

(5) --halogen,

(6) --C₁₋₃ alkyl,

(7) --S(O)₂ C₁₋₃ alkyl, and

(8) --C(O)R₆.

One class within this genus are the compounds of formula I wherein:

M is selected from a group consisting of:

C₂, C₃, or C₄ mono or di- substituted, the substitutent independentlyselected from:

(a) hydroxy,

(b) Cl or F,

(c) phenyl or mono or di-substituted phenyl, the substitutentsindependently selected from

(1) C₁₋₃ alkoxy,

(2) C₁₋₃ alkyl,

(3) halogen,

Rb is selected from the group consisting of

(a) C₁₋₃ alkyl, and

(b) hydrogen,

Rc is phenylC₁₋₄ alkyl wherein the phenyl is optionally mono, di- ortrisubstituted, the substitutents independently selected from

(1) halogen,

(2) trifluoromethyl,

(3) C₁₋₃ alkyl.

One subclass within this class are the compounds of formula I wherein:

R₁ is ##STR8## where B is (a) phenyl or mono, di or tri- substitutedphenyl wherein the substitutents are independently chloro, methyl,phenyl or CF₃ ;

(b) pyridyl or mono di or trisubstituted pyridyl wherein thesubstitutents are independently chloro, methyl, phenyl or CF₃ ; and

(c) thiophene or mono or disubstituted thiophene wherein thesubstitutents are independently chloro, methyl, phenyl or CF₃.

A second genus within the above described embodiment are the compoundsof formula I wherein; ##STR9## is optionally mono di or trisubstituted,wherein the group is selected from ##STR10## the optional substitutentsresiding at 1, 2, or 3 of the positions 1, 2, 3, 4, 5, 6, 7 and 8 of theabove groups, the substituents selected from the group consisting of

(1) hydroxy,

(2) cyano,

(3) --C(O)NR₆ R₇,

(4) --NR₆ R₇,

(5) --NR₆ COR₇,

(6) --halogen,

(7) --CF₃,

(8) --C₁₋₃ alkyl,

(9) --S(O)₂ C₁₋₃ alkyl.

As is clear from the examples and schemes, the designation: ##STR11## informula I is interchangable with (CH₂)_(l) or (CH₂)_(m) respectively. Asappreciated by those of skill in the art, halo as used herein areintended to include chloro, fluoro, bromo and iodo.

Exemplifying the invention are the compounds of the examples includingthe group consisting of

(a)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-1-methanesulfonyl-spiro[indoline-3,4'-piperidine];

(b)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-spiro[1H-indene-1,4'-piperidine];

(c)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]]-spiro[(3-indanone)-1,4'-piperidine];

(d)1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-phenyl-4-acetamido-piperidine;

(e)1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-benzyl-piperazine;

(f)1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-phenyl-piperidine].

(g)1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-(2-pyridyl)-piperazine;and

(h)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-1-acetyl-spiro[indoline-3,4'-piperidine].

Compounds of formula I within the scope of this invention are furtherdetailed in the claims appended to this application.

In an alternative embodiment may be co-administered with a β2-agonistsuch as Bambuterol, U.S. Pat. No. 4,419,364 issued to Draco on Dec. 6,1983; Bitolterol mesylate, U.S. Pat. No. 4,138,581 issued to SterlingFeb. 6, 1979; Carbuterol, U.S. Pat. No. 3,763,232 issued to Smith KlineOct. 2, 1973; Clenbuterol, U.S. Pat. No. 3,536,712 issued to BoehringerIngelheim Oct. 27, 1970; Dopexamine, U.S. Pat. No. 4,645,768 issued toFisons Feb. 24, 1987; Formoterol, U.S. Pat. No. 3,994,974 issued toYamanouchi Nov. 30, 1976; Mabuterol, U.S. Pat. No. 4,119,710 issued toBoehringer Ingelheim Oct. 10, 1978; Pirbuterol hydrochloride U.S. Pat.No. 3,700,681 issued to Pfizer Oct. 24, 1972; Procaterol hydrochlorideU.S. Pat. No. 4,026,897 issued to Otsuka May 31, 1977; Ritodrinehydrochloride U.S. Pat. No. 3,410,944 issued to North American PhilipsNov. 12, 1968; Brosaterol, U.S. Pat. No. 4,276,299 issued to Zambon Jun.30, 1981 and U.S. Pat. No. 4,520,200 issued to Zambon May 28, 1985;Cimaterol, U.S. 4,407,819 issued to American Cyanamid Oct. 4, 1983;Docarpamine, U.S. 4,228,183 issued to Tanabe Oct. 14, 1980; Salmeterol,U.S. Pat. No. 4,992,474 issued to Glaxo Feb. 21, 1991 and U.S. Pat. No.5,091,422 issued to Glaxo Feb. 25, 1992.

The compounds of formula I are particularly useful in the treatment ofdiseases or conditions that are advantageously treated by contomitantantagonism of both NK1 and NK3 receptors or NK1, NK2 and NK3 receptors.these diseases include neuropathy, such as diabetic or peripheralneuropathy and chemotherapy-induced neuropathy; asthma; osteoarthritis;rheumatoid arthritis; and migraine.

In a second alternative embodiment the compounds of formula I may beco-administered with another NK1 or NK2 antagonist such as thosedescribed in Appln No. DO-139125, filed 08-Jun-78, Pub. 12-Dec-79; ApplnNo. EP-82568, filed 22-Dec-81, Pub. 29-Jun-83; Appln No. EP-490379,filed 13-Dec-90, Pub. 17-Jun-92; Appln No. EP-353732, filed 05-Aug-88,Pub. 07-Feb-90; Appln No. EP-161007, filed 13-Jan-84, Pub. 13-Nov-85;Appln No. EP-385-43, filed 28-Feb-89, Pub. 05-Sep-90; Appln No.WO8301251, filed 09-Oct-81, Pub. 14-Apr-83; Appln No. BE-894602, filed09-Oct-81, Pub. 31-Jan-83; Appln No. DE3205991, filed 19-Feb-82, Pub.01-Sep-83; Appln No. EP-327009, filed 02-Feb-88, Pub. 09-Aug-89; ApplnNo. EP-336230, filed 05-Apr-88, Pub. 11-Oct-89; Appln No. 394989, filed28-Apr-89, Pub. 31-Oct-90; Appln No. AU9068010, filed 22-Dec-89, Pub.27-Jun-91; Appln No. EP-482539, filed 24-Oct-90, Pub. 29-Apr-92; ApplnNo. EP-443132, filed 10-Dec-90, Pub. 28-Aug-91; Appln No. EP-498069,filed 21-Dec-90, Pub. 12-Aug-92; Appln No. WO9222569, filed 19-Jun-91,Pub. 23-Dec-92; Appln No. JO4297492, filed 24-Oct-91, Pub. 21-Oct-92;Appln No. U.S. Pat. No. 4,997,853, filed 02-Dec-88, Pub. 05-Mar-91;Appln No. EP-272929, filed 24-Dec-86, Pub. 29-Jun-88; Appln No.EP-360390, filed 25-Jul-88, Pub. 28-Mar-90; Appln No. U.S. Pat. No.3,862,114, filed 22-Nov-71, Pub. 21-Jan-75; Appln No. EP-219258, filed30-Sep-85, Pub. 22-Apr-87, Appln No. U.S. Pat. No. 4,742,156, filed30-Sep-85, Pub. 03-May-88; Appln No. EP-401177, filed 29-May-89, Pub.05-Dec-90; Appln No. WO9202546, filed 03-Aug-90, Pub. 20-Feb-92; ApplnNo. EP-176436, filed 26-Sep-84, Pub. 02-Apr-86; Appln No. U.S. Pat. No.4,680,283, filed 26-Sep-84, Pub. 14-Jul-87; Appln No. WO9220661, filed22-May-91, Pub. 26-Nov-92; Appln No. EP-520555, filed 24-Jun-91, Pub.30-Dec-92; Appln No. EP-347802, filed 20-Jun-88, Pub. 27-Dec-89; ApplnNo. EP-412542, filed 10-Aug-89, Pub. 13-Feb-91; Appln No. WO9005729,filed 23-Nov-88, Pub. 31-May-90; Appln No. WO9005525, filed 23-Nov-88,Pub. 31-May-90; Appln No. EP-436334, filed 04-Jan-90, Pub. 10-Jul-91;Appln No. WO9118878, filed 31 -May-90, Pub. 12-Dec-91; Appln No.WO9118899, filed 01 -Jun-90, Pub. 12-Dec-91; Appln No. WO9201688, filed23-Jul-90, Pub. 06-Feb-92; Appln No. WO9206079, filed 28-Sep-90, Pub.16-Apr-92; Appln No. WO9212152, filed 03-Jan-91, Pub. 23-Jul-92; ApplnNo. WO9212151, filed 10-Jan-91, Pub. 23-Jul-92; WO9215585, filed01-Mar-91, Pub. 29-Apr-92; Appln No. WO022-676, filed 22-May-91, Pub.26-Nov-92; Appln No. WO9221677, filed 3 1-May-91, Pub. 10-Dec-92; ApplnNo. WO9300331, filed 20-Jun-91, Pub. 07-Jun-93; Appln No. WO9300330,filed 21-Jun-91, Pub. 07-Jan-93; Appln No. WO9109844, filed 11-Jul-91,Pub. 11 -Jul-91; Appln No. EP-429366, filed 23-Nov-89, Pub. 29-May-91;Appln No. EP-43077 1, filed 23-Nov-89, Pub. 05-Jun-91; Appln No.EP-514274, filed 17-May-91, Pub. 19-Nov-92; Appln No. EP-514276, filed17-May-91, Pub. 19-Nov-92; Appln No. EP-514275, filed 17-May-91, Pub.19-Nov-92; Appln No. EP-514273, filed 17-May-91, Pub. 19-Nov-92; ApplnNo. EP-428434, filed 06-Nov-89, Pub. 22-May-91; Appln No. EP-474561,filed 09-May-90, Pub. 11-Mar-92; Appln No. EP-512901, filed 03-May-91,Pub. 1 1-Nov-92; Appln No. EP-512902, filed 03-May-91, Pub. 1 1-Nov-92;Appln No. EP-515240, filed 03-May-91, Pub. 25-Nov-92; Appln No.US4472305, filed 17-May-83, Pub. 18-Sep-84; Appln No. US4839465, filed20-Jan-87, Pub. 13-Jun-89; Appln No. EP-101929, filed 28-Jul-82, Pub.07-Mar-84; Appln No. WO9102745, filed 16-Aug-89, Pub. 07-Mar-91; ApplnNo. U.S. Pat. No. 3,912,711, filed 03-Jul-72, Pub. 14-Oct-75; Appln No.U.S. Pat. No. 4,059,693, filed 11-Jun-76, Pub. 22-Nov-77; Appln No. U.S.Pat. No. 4,481,139, filed 13-Apr-83, Pub. 06-Nov-84; Appln No. U.S. Pat.No. 7,358,073, filed 24-Oct-88, Pub. 19-Dec-89; Appln No. U.S. Pat. No.7,261,627, filed 24-Oct-88, Pub. 07-Mar-89, which are herebyincorporated by reference.

As may be appreciated by those of skill in the neurokinin art, that itis generally important to treat a patient with a neurokinin mediateddisease, with an antagonist that is specific for the neurokininreceptors(s) mediating the disease. Thus a patient suffering from any ofthe diseases mediated by neurokinin-1 receptors, is generally mostadvantageously treated with a compound that is a potent antagonist ofneurokinin-1, but is at most a weak antagonist of neurokinin-3. Evenwhere antagonism of both neurokinin-1 and neurokinin-2 is desired, it isgenerally advantageous to avoid significant antagonism of neurokinin-3.Similarly, a patient suffering from any of the diseases mediated byneurokinin-3, is generally most advantageously treated with a compoundthat is a potent antagonist of neurokinin-3, but is at most a weakantagonist of neurokinin-1.

Accordingly, in a third embodiment the compounds of formula I are usefulas a diagnostic tool for assessing the degree to which neurokinin-3receptor activity (normal, overactivity or underactivity) is implicatedin a patient's symptoms. In this regard a compound of formula I is usedas an antagonist of the activity, for example including but notrestricted to tachykinin agonist-induced inositol phosphate turnover orelectrophysiological activation, of a cell sample obtained from apatient. Comparison of such activity in the presence or absence of acompound of formula I will disclose the degree of NK-3 receptorinvolvement in the mediation of agonist effects in that tissue.

Similarly, in a fourth embodiment, the compounds of formula I are usefulin assessing the degree to which a selected pharmacological effect, asmeasured either in vitro or in vivo, is due to activation of the NK-3receptor. In this instance the assays are performed in the presence orabsence of a compound of formula I, and comparison of the results willdisclose the degree of NK-3 receptor involvement.

The compounds of formula I are useful in the prevention and treatment ofa wide variety of clinical conditions (as detailed in thisspecification) which are characterized by overstimulation of thetachykinin receptors, in particular NK1, NK2 and NK3.

These conditions may include disorders of the central nervous systemsuch as anxiety, depression, psychosis and schizophrenia;neurodegenerative disorders such as AIDS related dementia, seniledementia of the Alzheimer type, Alzheimer's disease and Down's syndrome;demyelinating diseases such as multiple sclerosis and amyotrophiclateral sclerosis and other neuropathological disorders such as diabeticor peripheral neuropathy, AIDS related neuropathy, chemotherapy-inducedneuropathy, and neuralgia; respiratory diseases such as chronicobstructive airways disease, bronchopneumonia, bronchospasm and asthma;inflammatory diseases such as inflammatory bowel disease, psoriasis,fibrositis, osteoarthritis and rheumatoid arthritis; allergies such aseczema and rhinitis; hypersensitivity disorders such as poison ivy;ophthalmic diseases such as conjunctivitis, vernal conjunctivitis, andthe like; cutaneous diseases such as contact dermatitis, atopicdermatitis, urticaria, and other eczematoid dermatitis; addictiondisorders such as alcholism; stress related somatic disorders; reflexsympathetic dystrophy such as shoulder/hand syndrome; dysthymicdisorders; adverse immunological reactions such as rejection oftransplanted tissues and disorders related to immune enhancement orsuppression such as systemic lupus erythematosis; gastrointestinal (GI)disorders and diseases of the GI tract such as disorders associated withthe neuronal control of viscera such as ulcerative colitis, Crohn'sdisease and incontinence; disorders of bladder function; fibrosing andcollagen diseases such as scleroderma and eosinophilic fascioliasis;disorders of blood flow caused by vasodilation and vasospastic diseasessuch as angina, migraine and Reynaud's disease; and pain or nociception,for example, that is attributable to or associated with any of theforegoing conditions especially the transmission of pain in migraine.Hence, these compounds are readily adapted to therapeutic use for thetreatment of physiological disorders associated with the overstimulationof the tachykinin receptors, in particular NK1, NK2 and NK3.

The compounds of the present invention are particularly useful in thetreatment of pain or nociception and/or intimation and disordersassociated therewith such as, for example: neuropathy, such as diabeticor peripheral neuropathy and chemotherapy-induced neuropathy; asthma;osteoarthritis; rheumatoid arthritis; and migraine.

For the treatment of any of these diseases compounds of Formula I may beadministered orally, topically, parenterally, ICV, by inhalation sprayor rectally in dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carders, adjuvants and vehicles.The term parenteral as used herein includes subcutaneous injections,intravenous, intramuscular, intracistemal injection or infusiontechniques. In addition to the treatment of warm-blooded animals such asmice, rats, horses, cattle, sheep, dogs, cats, etc., the compounds ofthe invention are effective in the treatment of humans.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example starch, gelatin or acacia, and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets may be uncoated or they may be coated by known techniques todelay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distearatemay be employed. They may also be coated by the techniques described inthe U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotictherapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl- pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally- occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents. The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of formula I may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable non-irritatingexcipient which is solid at ordinary temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials are cocoa butter and polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compounds of Formula I are employed. (For purposesof this application, topical application shall include mouth washes andgargles.)

In the treatment of a condition associated with an excess oftachykinins, an appropriate dosage level will generally be about 0.001to 50 mg per kg patient body weight per day which can be administered insingle or multiple doses. Preferably, the dosage level will be about0.01 to about 25 mg/kg per day; more preferably about 0.05 to about 10mg/kg per day. A suitable dosage level may be about 0.00 1 to 25 mg/kgper day, about 0.005 to 10 mg/kg per day, or about 0.005 to 5 mg/kg perday. Within this range the dosage may be 0.005 to 0.05, 0.05 to 0.5 or0.5 to 5.0 mg/kg per day. The compounds may be administered on a regimenof 1 to 4 times per day, preferably once or twice per day.

TACHYKININ ANTAGONISM ASSAY

The compounds of this invention are useful for antagonizing tachykinins,in particular substance P and neurokinin A in the treatment ofgastrointestinal disorders, central nervous system disorders,inflammatory diseases, pain or migraine and asthma in a mammal in needof such treatment. This activity can be demonstrated by the followingassay.

A. Receptor Expression in COS

To express the cloned human neurokinin-1 receptor (NK1R) transiently inCOS, the cDNA for the human NK1R was cloned into the expression vectorpCDM9 which was derived from pCDM8 (INVITROGEN) by inserting theampicillin resistance gene (nucleotide 1973 to 2964 from BLUESCRIPT SK+)into the Sac II site. Transfection of 20 μg of the plasmid DNA into 10million COS cells was achieved by electroporation in 800 μl oftransfection buffer (135 mM NaCl, 1.2 mM CaCl₂, 1.2 mM MgCl₂, 2.4 mM K₂HPO₄, 0.6 mM KH₂ PO₄, 10 mM glucose, 10 mM HEPES pH 7.4) at 260 V and950 uF using the IBI GENEZAPPER (IBI, New Haven, Conn.). The cells wereincubated in 10% fetal calf serum, 2 mM glutamine, 100U/mlpenicillin-streptomycin, and 90% DMEM media (GIBCO, Grand Island, N.Y.)in 5% CO₂ at 37 ° C. for three days before the binding assay.

Similar methods were used to express the human NK2 and NK3 receptors.

B. Stable Expression in CHO

To establish a stable cell line expressing the cloned human NK1R, thecDNA was subcloned into the vector pRcCMV (INVITROGEN). Transfection of20 μg of the plasmid DNA into CHO cells was achieved by electroporationin 800 μl of transfection buffer suplemented with 0.625 mg/ml Herringsperm DNA at 300 V and 950 uF using the IBI GENEZAPPER (IBI). Thetransfected cells were incubated in CHO media [10% fetal calf serum, 100U/ml pennicilin-streptomycin, 2 mM glutamine, 1/500hypoxanthine-thymidine (ATCC), 90% IMDM media (JRH BIOSCIENCES, Lenexa,Kans.), 0.7 mg/ml G418 (GIBCO)] in 5% CO₂ at 37° C. until colonies werevisible. Each colony was separated and propagated. The cell clone withthe highest number of human NK1R was selected for subsequentapplications such as drug screening.

Similar methods were used to express the human NK2 and NK3 receptors.

C. Assay Protocol using COS or CHO

The binding assay of human NK1R expressed in either COS or CHO cells isbased on the use of ¹²⁵ I-substance P (¹²⁵ I-SP, from DU PONT, Boston,Mass.) as a radioactively labeled ligand which competes with unlabeledsubstance P or any other ligand for binding to the human NK1R. Monolayercell cultures of COS or CHO were dissociated by the non-enzymaticsolution (SPECIALTY MEDIA, Lavallette, N.J.) and resuspended inappropriate volume of the binding buffer (50 mM Tris pH 7.5, 5 mM MnCl₂,150 mM NACl, 0.04 mg/ml bacitracin, 0.004 mg/ml leupeptin, 0.02 mg/mlBSA, 0.01 mM phosphoramidon) such that 200 μl of the cell suspensionwould give rise to about 10,000 cpm of specific ¹²⁵ I-SP binding(approximately 50,000 to 200,000 cells). In the binding assay, 500 ul ofcells were added to a tube containing 20 μl of 1.5 to 0.25 nM of ¹²⁵I-SP and 5 μl of unlabeled substance p or any other test compound inDMSO. The tubes were incubated at 4° C. or at room temperature for 1hour with gentle shaking. The bound radioactivity was separated fromunbound radioactivity by GF/C filter (BRANDEL, Gaithersburg, Md.) whichwas pre-wetted with 0.1% polyethylenimine. The filter was washed with 3ml of wash buffer (50 mM Tris pH 7.5, 5 mM MnCl₂, 150 mM NaCl) threetimes and its radioactivity was determined by gamma counter. A similarassay was used for NK2 and NK3 except ¹²⁵ I-NKA or ¹²⁵ I-methyl-Phe⁷-NKB were used as the ligand.

The activation of phospholipase C by NK1R may also be measured in CHOcells expressing the human NK1R by determining the accumulation ofinositol monophosphate which is a degradation product of IP₃. CHO cellsare seeded in 12-well plate at 250,000 cells per well. After incubatingin CHO media for 4 days, cells are loaded with 0.025 uCi/ml of ³H-myoinositol by overnight incubation. The extracellular radioactivityis removed by washing with phosphate buffered saline. LiCl is added tothe well at final concentration of 0.1 mM with or without the testcompound, and incubation is continued at 37° C. for 15 min. Substance Pis added to the well at final concentration of 0.3 nM to activate thehuman NK1R. After 30 min of incubation at 37° C., the media is removedand 0.1N HCl is added. Each well is sonicated at 4° C. and extractedwith CHCl₃ /methanol (1:1). The aqueous phase is applied to a 1 ml DowexAG 1×8 ion exchange column. The column is washed with 0.1N formic acidfollowed by 0.025M ammonium formate-0.1N formic acid. The inositolmonophosphate is eluted with 0.2M ammonium formate-0.1N formic acid andquantitated by beta counter. similar methods were used to assessantagonism at the NK2 and NK3 receptors, except NKA or NKB were used asthe stimulating agonists.

The compounds of of Formula I as Exemplified in the EXAMPLES below havebeen found to displace radioactive ligand for the NK-1 receptor at aconcentration range of 0.01 nM to 1.0 μM, for the NK-2 receptor, 0.01 nMto 5 μM, and for the NK-3 receptor, 1.0 nM to 10 μM. Further data isprovided in Table 1.

                  TABLE I                                                         ______________________________________                                         ##STR12##                                                                     ##STR13##        NK.sub.1 (nM)                                                                          NK.sub.2 (nM)                                                                           NK.sub.3 (nM)                            ______________________________________                                         ##STR14##       250       45       1.5                                        ##STR15##       100      200       8                                          ##STR16##       500      200       8                                          ##STR17##                 30       0.75                                       ##STR18##                 30       0.6                                        ##STR19##                250       3                                          ##STR20##                220       4                                          ##STR21##                 40       1                                         ______________________________________                                    

Several methods for preparing the compounds of this invention areillustrated in the following Schemes and Examples.

The compounds of the present invention are prepared by alkylatingazacycle 1, in which R₁ =H, under appropriate conditions (Scheme 1). Therequired azacycle staging materials are prepared using methods describedin the literature; more specifically, as described in Ong, H. H. et al,Journal of Medicinal Chemistry, 1983,26, 981-986, and Nargund, R. et al,Merck patent application no. 18899. None of the compounds in thesereferences are claimed to be neurokinin antagonists.

Thus, azacycle 1 (R₁ =H) is combined with the appropriate aldehyde andthe intermediate imine is reduced to the tertiary amine chemically (e.g.using sodium cyanoborohydride) or catalytically (e.g. using hydrogen andpalladium on carbon or Raney nickel catalyst) (Scheme 1). The aldehydeneeded for this reaction can be prepared by methods generally known inthe chemical literature; for the purposes of the present invention thepreparation of a representative aldehyde is described in Hale, J. J.;Finke, P. E.; MacGoss, M. Bioorganic and Medicinal Chemistry Letters, 2,(Feb. 1993).

In an alternative embodiment of the present invention, azacycle 1 (R₁=H) can be alkylated with an alkyl halide or alkyl sulfonate ester (withor without an added base to neutralize the mineral acid or sulfonic acidby-product) to give the desired compound (Scheme 1). The alkyl halide oralkyl sulfonate needed for this reaction can be prepared by methodsgenerally known in the chemical literature; for the purposes of thepresent invention an aldehyde, prepared as described above, can bereduced to an alcohol with sodium borohydride, diisobutylaluminumhydride or lithium aluminum hydride, and the product alcohol convened toeither the alkyl halide using methods described in March J. "AdvancedOrganic Chemistry", 3rd ed., John Wiley & Sons, New York, pp. 382-384(1985), or alkyl sulfonate ester using methods described in March J."Advanced Organic Chemistry", 3rd ed., John Wiley & Sons, New York, p.444 (1985).

In an alternative embodiment of the present invention, 1 (R₁ =H) can beacylated to give the tertiary amide and subsequent reduction with astrong reducing agent (e.g. diborane including borane dimethylsulfide;and, lithium aluminum hydride) will give the desired compound (Scheme1). The acylating agent needed for this reaction can be prepared bymethods generally known in the chemical literature; for the purposes ofthe present invention an aldehyde, prepared as described above, can beoxidized using such commonly used reagents as permanganate in acid orsilver oxide, and the resulting acid activated as an acid chloride ormixed anhydride which can be used to acylate 1 (R₁ =H). The productamide can in and of itself be a neurokinin antagonist or can be reducedwith a strong reducing agent, such as diborane of lithium aluminumhydride, to give the tertiary amine.

Optionally, compound 1 formed in the alkylation step may be furthermodified in subsequent reactions. In one illustration of such anapproach,the aldehyde fragment contained a t-butoxycarbonylamino group(Example 2). After reductive amination, the t-butoxycarbonyl protectinggroup is removed by treatment with a strong acid such as trifluoroaceticacid or formic acid and the resulting amine is acylated to furnish thedesired compounds (Example 3). Alternatively, the protecting group mayalso be present in the azacycle portion as illustrated with abenzyloxycarbonyl group in Example 6. Thus an azacycle containing abenzyloxycarbonylindoline (prepared in example 4) is alkylated with analdehyde in the presence of a reducing agent. Next, the protecting groupis removed to liberate a free amine (example 7) and the amine is furtherreacted to provide additional analogs (Example 8). ##STR22##

EXAMPLE 1

3-(S)-(3,4-Dichlorophenyl)-4-(N-benzyl-N-methylamino)-4-oxo-butanal.

A solution of 4.8 g (19.6 mmol) of2-(S)-(3,4-dichlorophenyl)-4-pentenoic acid in 50 mL of CH₂ Cl₂ and 10drops of DMF was cooled in an ice bath and 1.8 mL of oxalyl chloride wasadded. The solution was stirred for 15 min after gas evolution hadceased. The mixture was concentrated in vacuo and the residue wasdiluted with 50 mL of CH₂ Cl₂. The solution was cooled in ice bath as2.6 mL (20.4 mmol) of N-benzyl-methyl amine and 1.7 mL (21 mmol) ofpyridine were added. After stirring for 1 h at ice bath temperature thesolution was diluted with CH₂ Cl₂ and washed with water, 1.2N HCl andbrine. The solution was dried over Na₂ SO₄ and concentrated to giveN-benzyl-N-methyl-2(S)-(3,4-dichlorophenyl)-4-pentenoamide as an oil.

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 2.4 (m, 1H); 2.8 (m, 1H); 2.82 and 2.95 (2 s, 3H);3.52-3.8 (m, 1H); 4.32 and 4.6 (ABq) and 4.57 (s, 2H); 5.0(m, 2H), 5.7(m, 1H), 6.9-7.4 (m, 8H).

The amide obtained above was dissolved in 40 mL of acetone, 20 mL oft-butanol and 20 mL of water. To this solution 5 mg of Osmium tetroxideand 2.5 g (21.4 mmol) of 4-methylmorpholine N-oxide were added. Afterstirring overnight, the reaction was quenched with approximately 2 g ofNa₂ SO₃ and concentrated to 25% of the original volume. The residue waspartitioned between water and 1:1 ether (Et₂ O), ethyl acetate (EtOAc),the layers were separated and the aqueous layer was extracted with Et₂O:EtOAc. Each organic layer was washed with water, brine and dried byfiltering through Na₂ SO₄. The filtrate was concentrated to afford thecrude diol.

A solution of the diol in 60 mL of tetrahydrofuran (THF) and 20 mL ofwater was treated with 4.5 g (21 mmol) of Sodium periodate. Afterstirring for 2 h, the reaction was diluted with Et₂ O:EtOAC and washedwith water and brine. The organic layer was dried (Na₂ SO₄ ) and thefiltrate was concentrated. The residue was purified by flash columnchromatography using a gradient of 30-50% EtOAC/hexane to furnish 3.7 g(54% yield for three steps) of the title compound as a thick oil.

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 2.65 (m, 1H); 2.85 and 2.91 (2 s, 3H); 3.54 (m, 1H);4.25-4.65 (m, 3H), 7.0-7.4 (m, 8H); 9.75 and 9.78 (2 s, 1H).

Mass Spectrum (CI) 351 (³⁷ Cl+³⁵ Cl isotope), 349 (³⁵ Cl+³⁵ Cl isotope).

EXAMPLE 2

1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-1-methanesulfonyl-spiro[indoline-3,4'-piperidine].

To a solution of 0.36 g (1.03 mmol) of3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butanal(Example 1) in 3 mL of methanol were added 0.32 g (1.05 mmol) of1-methanesoufonyl-spiroindoline-3,4'-piperidine hydrochloride and 0.3 gof powedered 4 Å molecular sieves. After 1 h, 1N NaCNBH₃ in THF (3 mL)was added over 10 min with a syringe pump. Afeter 1 h, when the reactionwas complete by TLC, the mixture was filtered through a pad of celite,the reaction flask and the pad were rinsed with methanol. The filtratewas concentrated to approximately 5 ml and the residue was partitionedbetween saturated NaHCO₃ and Et₂ O:EtOAC. The organic layer was washedwith water, brine and dried over NA₂ SO₄. The filtrate was concentratedand the residue was chromatographed on a flash column using 49:49:2EtOAc: Hexane: triethylamine to furnish 0.52 g (84%) of the titlecompound as a foam.

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.5-2.9 (m, 12 H), 2.87 & 2.88 (2 s, 3 H), 2.88 & 2.98 (2s, 3 H), 3.73 & 3.76 (2 s, 2H), 3.9 (m, 1H), 4.3-4.7 (m, 2H), 7.0-7.5(m, 12 H).

Mass Spectrum (CI) 602 (³⁷ Cl+³⁵ Cl isotope), 600 (³⁵ Cl+³⁵ Cl isotope).

The following compounds were prepared by reacting the appropriate aminewith 3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butanal(Example 1) according to the procedure of Example 2.

EXAMPLE 3

1'[3-(S )-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-spiro[1H-indene-1,4'-piperidine]

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.2-3.0 (m, 12 H), 2.91 & 2.99 (2 s, 3 H), 3.9 (m, 1H),4.4-4.7 (m, 2 H), 6.7 (m, 2H), 7.0-7.5 (m, 12 H).

Mass Spectrum (CI) 521 (³⁷ Cl+³⁵ Cl isotope), 519 (³⁵ Cl+³⁵ Cl isotope).

EXAMPLE 4

1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-spiro[(3-indanone)-1,4'-piperidine].

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.4-3 (m, 12 H), 2.51 & 2.54 (2 s, 2 H), 2.89 & 2.98 (2 s,3 H), 3.9 (m, 1H), 4.3-4.7 (m, 2H), 7.05-7.75 (m, 12 H).

Mass Spectrum (CI) 537 (³⁷ Cl+³⁵ Cl isotope), 535 (³⁵ Cl+³⁵ Cl isotope).

EXAMPLE 5

1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-phenyl-4-acetamido-piperidine.

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.5-2.8 (m, 12 H), 1.97 & 1.98 (2 s, 3 H), 2.87 & 2.97 (2s, 3 H), 3.9 (m, 1 H), 4.3-4.7 (m, 2 H), 5.5 (br d, 1 H), 7.0-7.45 (m,13 H).

Mass Spectrum (CI) 554 (³⁷ Cl+³⁵ Cl isotope), 552 (³⁵ Cl+³⁵ Cl isotope).

EXAMPLE 6

1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-benzyl-piperazine.

To a solution of 0.12 g (0.34 mmol) of3-(S)-(3,4-Dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butanal(Example 1) in 2 mL of methanol were added 57 μL (0.33 mmol) ofN-benzyl-piperazine, 18 μL (0.31 mmol) of acetic acid and 0.2 g ofpowedered 4 Å molecular sieves. After 1 h, 1N NaCNBH3 in THF (0.8 mL)was added over 10 min with a syringe pump. After 1 h, when the reactionwas complete by TLC, the mixture was filtered through a pad of celite,the reaction flask and the pad were rinsed with methanol. The filtratewas partitioned between saturated NaHCO3 and Et₂ O:EtOAC. The organiclayer was washed with water, brine and dried over NA₂ SO₄. The filtratewas concentrated and the residue was purified by prep TLC using 2%Et3N/EtOAc to furnish 97 mg (58%) of the title compound as a foam.

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.5-2.8 (m, 12 H), 2.86 & 2.95 (2 s, 3 H), 3.46 & 3.48 (2s, 2 H), 3.9 (m, 1 H), 4.3-4.7 (m, 2 H), 7.0-7.5 (m, 13 H).

Mass Spectrum (CI) 512 (³⁷ Cl+³⁵ Cl isotope), 510 (³⁵ Cl+³⁵ Cl isotope).

The following compounds were synthesized by substituting the requiredamine for N-benzyl-piperazine in example 6.

EXAMPLE 7

1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-phenyl-piperidine].

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.5-3.0 (m, 12 H), 2.89 & 2.97 (2 s, 3 H), 3.4-4.1 (m, 1H), 3.95 (m, 1 H), 4.3-4.7 (m, 2 H), 7.0-7.5 (m, 13 H).

Mass Spectrum (CI) 497 (³⁷ Cl+³⁵ Cl isotope), 495 (³⁵ Cl+³⁵ Cl isotope).

EXAMPLE 8

1-[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-4-(2-pyridyl)-piperazine.

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.6 (br, 1 H), 1.85 (m, 1 H), 2.2-2.6 (m, 6 H), 2.87 &2.97 (2 s, 3 H), 3.46 (m, 4 H), 3.9 & 4.0 (2 t, 1 H, J=7 Hz), 4.3-4.7(m, 2 H) 6.61 (m, 2 H), 7.0-7.5 (m, 9 H), 8.16 (m, 1 H).

Mass Spectrum (CI) 499 (37Cl+³⁵ Cl isotope), 497 (³⁵ Cl+³⁵ Cl isotope).

EXAMPLE 9

1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-1-acetyl-spiro[indoline-3,4'-piperidine].

¹ H NMR (CDCl₃, ppm ranges are given because of amide rotomers and linebroadening) δ 1.5-3.0 (m, 12 H), 2.22 & 2.23 (2 s, 3 H), 2.89 & 2.98 (2s, 3 H), 3.78 & 3.81 (2 s, 2 H), 3.9 (m, 1 H), 4.3-4.7 (m, 2 H), 7.0-7.5(m, 11 H), 8.18 (d, 1 H, J=8 Hz).

Mass Spectrum (CI) 566 (37Cl+³⁵ Cl isotope), 564 (³⁵ Cl+³⁵ Cl isotope).

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 3                                                  (2) INFORMATION FOR SEQ ID NO:1:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 11 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (iii) HYPOTHETICAL: NO                                                        (iv) ANIT-SENSE: NO                                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                       MetLeuGlyPhePheGlnGlnProLysProArg                                             1510                                                                          (2) INFORMATION FOR SEQ ID NO:2:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                         (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                       MetLeuGlyValPheSerAspThrLysHis                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:3:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                     (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                       GlyProTyrTrpGlyHisSerTrpHisXaa                                                1510                                                                      

What is claimed is:
 1. A compound of Formula Ia ##STR23## wherein thegroup ##STR24## is selected from the group consisting of ##STR25## thesubstituents residing at 1 or 3 of the positions 1,3,4,5,or 6 of theabove groups the substituents selected from the group consisting of:(1)hydroxy, (2) oxo, (3) cyano, (4) --NHR₆, (5) --NR₆ R₇, (6) --NR₆ COR₇,(7) --halogen, (8) --CF₃, (9) --C₁₋₃ alkyl, (10) --S(O)₂ C₁₋₃alkylwherein R₆ and R₇ are hydrogen or C₁₋₃ alkyl, or mono- ordi-substituted C₁₋₃ alkyl, the substituents independently selected from:(1) C₁₋₃ alkoxy, (2) hydroxy, (3) oxo, (4) halogen; and R₁ is ##STR26##where B is selected from: (a) phenyl or mono- or di-substituted phenylwherein the substituents are independently chloro, methyl, phenyl or CF₃;(b) pyridyl or mono-, di- or tri-substituted pyridyl wherein thesubstituents are independently chloro, methyl, phenyl or CF₃ ; and (c)thiophene or mono- or di-substituted thiophene wherein the substitutentsare independently chloro, methyl, phenyl or CF₃.
 2. A compound selectedfrom the group consisting of:(a)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-1-methanesulfonyl-spiro[indoline-3,4'-piperidine];(b)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-spiro[1H-indene-1,4'-piperidine];(c)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-spiro[(3-indanone)-1,4'-piperidine];(d)1'[3-(S)-(3,4-dichlorophenyl)-4-(N-benzyl-N-methyl)amino-4-oxo-butyl]-1-acetyl-spiro[indoline-3,4'-piperidine].3. A compound according to claim 1 wherein B is phenyl or mono or di-substituted phenyl wherein the substitutents are independently chloro,methyl, phenyl or CF₃.
 4. A method of treating or preventing asthma in apateint in need thereof which comprises the administration to thepatient of a non-toxic therapeutically effective amount of the compoundof claim 1.